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中国农学通报

中国农学通报 ›› 2011, Vol. 27 ›› Issue (9): 347-351.

所属专题： 生物技术

• 生物技术科学 • 上一篇    下一篇

紫苏苯丙氨酸解氨酶基因片段克隆及序列分析

吕晓玲 孙雪梅 王芳 郝磊 孙晶磊   

  • 收稿日期:2010-12-28 修回日期:2011-01-30 出版日期:2011-04-25 发布日期:2011-04-25

Molecular Cloning and Sequence Analysis of Phenylalanine Ammonia-lyase Gene Fragment in Perilla frutescens

  • Received:2010-12-28 Revised:2011-01-30 Online:2011-04-25 Published:2011-04-25

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摘要：

苯丙氨酸解氨酶（Phenylalanine ammonia-lyase，PAL）是迷迭香酸合成途径中苯丙氨酸支路的关键酶，它催化苯丙氨酸生成肉桂酸，完成该支路第一步反应。本实验利用同源克隆方法成功克隆了紫苏PAL基因cDNA片段，命名为PerPAL-1（GenBank登录号：HQ388347.1），该片段长399 bp，编码133个氨基酸。通过氨基酸序列比对分析，发现其氨基酸序列与丹参和藿香PAL该片段的同源性分别高达96%和95%。PAL系统进化树表明PerPAL-1与唇形科植物的PAL亲缘关系最近。PerPAL-1基因在叶中表达最强，根中次之，而在茎中表达最弱。

关键词: 两步法RT-PCR, 两步法RT-PCR

Abstract:

Phenylalanine ammonia-lyase (PAL), responsible for catalyzing the conversion of phenylalanine to cinnamic acid to finish the first step of phenylalanine pathway, was the key enzyme during the biosynthesis of rosmarinic acid. The cDNA fragment of PAL gene was successfully cloned by homology cloning method (Accession No. HQ388347.1), 399 bp and encoded 133 amino acids. It was designated as PerPAL-1. The results of amino acid sequence analysis showed that the identity of the sequence of PerPAL-1 amino acid with that of Salvia miltiorrhiza and Agastache rugosa was 96% and 95%, respectively. Phylogenetic tree analysis revealed that PerPAL-1 had closer relationship with PALs from Lamiaceae plants than those of other plants. The expression of PerPAL-1 gene was the strongest in young leaves and the weakest in stems.

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